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Effects of PD1 and CTLA4 Inhibition on Cytotoxicity of CD8+ T-Cells Targeting Ewing’s Sarcoma Cell Line A673
No full textPD1 and CTLA4 inhibitors are often used to improve the effectiveness of TCR immunotherapy. However, their efficacy varies across tumor types. The aim of this research was to determine if PD1 and CTLA4 inhibition improves the cytotoxicity of primary CD8+ T-cells transfected with genes for a TCR targeting the Ewing’s Sarcoma cell line A673. For each experiment that was run, two co-cultures were established: A673 cells co-cultured with non-transfected (NT) CD8+ T-cells and A673 cells co-cultured with transfected CD8+ T-cells. One set of triplicates from each co-culture type had Anti-PDL1 (5ug/ml) and Anti-CTLA4 (5ug/ml) while the other set of triplicates had no added antibodies. The first assay ran was a fluorescence microscopy assay in which PI was added to all wells to identify dead cells and pictures were taken of the co-cultures every hour for 72 hours. The data was quantified using Cell Profiler. However the data had very high standard deviations so a flow cytometry assay was used to verify the results. For this assay the cells were removed from co-culture after 48 hours, stained using PI and Annexin V, then analyzed using flow cytometry. A 4% increase in cytotoxicity was observed. The results showed that Anti-PD1 and Anti-CTLA4 treatments increase cytotoxicity of CD8+ T-cells against A673 cells in vitro over a short period of time. This increase in such a short time frame, although slight, indicates that further research on the effects over a larger time frame and in vivo could be worthwhile
Microwave Spectrum Analysis of 5-chloro-2-fluorophenol
No full textThe purpose of microwave spectroscopy is to examine the change of rotation of a molecule to understand its’ shape, and how it would interact with other compounds. In our research, the experimental microwave spectrum of 5-chloro-2-fluorophenol was measured in an 8-18 GHz range. The spectrum was measured in 10 averages in a 10,000 MHz range. The calculated spectrum was obtained using Gaussian 03W software set at OPT+Freq anharmonic with basis set at 6-311 G+ (d,p). The calculated spectrum was compared to the measured spectrum. Broadband Analysis Software, was used for initial fit. More precise fit was used with AABS software. The rotational, distortion, and nuclear quadrupole coupling constants were observed. Rotational constants relate to shape. Distortion constants relate to how the compound stretches as it moves. Nuclear quadrupole coupling constants were observed as the nucleus of the chlorine on the fifth position of the carbon ring split the spectrum. The value of the nuclear quadrupole coupling constants indicated how skewed the spectrum was. Comparison of the measured experimental constants and calculated constants showed the cis conformer of 5-chloro-2-fluorophenol was likely measured
Investigating the Optimal Time to Harvest Actinomyces Species for Mouse Inoculations
No full textWithin the last few years, more states have been legalizing the use of cannabis to treat certain diseases. Though the drug has been effective, it is still extremely addictive, and the overuse of the drug can affect the general makeup of our oral microbiomes. Oral microbiomes are the communities of bacteria that live within our mouths and when their structure is changed, they may affect our cognitive system. One specific species of bacteria, Actinomyces, had a significant increase in abundance in the mouth of chronic cannabis smokers. To test the effects of the bacteria, the bacteria was regularly inoculated into the oral captives of mice. However, the bacteria refused to colonize within the mice, and without colonization, no effects can be observed. Thus, our research aimed to troubleshoot potential reasons as to why the bacteria refused to colonize. A growth curve of Actinomyces meyeri and Actinomyces odontolyticus was conducted to discover the phase of the bacteria, which could be a potential reason for it not colonizing. The creation of the growth curve involved growing the bacteria for 48 hours, inserting the samples in an absorbance reader, and using GraphPad Prism to create the growth curve from the data collected. From the data, we were able to conclude that the phase of the bacteria was indeed a factor for it not colonizing. We were also able to determine an optimum harvesting time for the bacteria. These findings will assist in future research that involve the use of the Actinomyces species
Standardizing an Audio-Visual Database of Misophonic Triggers
No full textMisophonia is a disorder of decreased tolerance to specific sounds or their associated stimuli. These sounds (e.g., chewing or scratching), also known as misophonic triggers, cause emotional distress, such as extreme irritation, anger, disgust, or anxiety to people with this condition. To study the mechanism underlying misophonic responses, specific trigger stimuli is required to evoke responses/reactions from participants in lab environments; however, with such high diversity in trigger categories/sources and the time-consuming nature of data organization, few attempts have been made at making a trigger database. Furthermore, past literature mainly focused on studying the acoustic aspect of misophonia, disregarding the visual association component of triggers. Our lab aims to create a public audio-visual database with diverse triggers. We are developing this goal by standardizing the video files and altering their viability and physical properties. One must understand what is happening in the video, and the trigger in the video must potentially provoke someone with misophonia. Our audio-visual database of triggers will aid researchers studying misophonia and help those with misophonia looking to discover more about their triggers
The Effect of Exposure to Thuricide on DNA methylation patterns of Fragaria ananassa
No full textThe purpose of this study was to analyze how exposing Fragaria ananassa DNA to Thuricide for different periods of time would impact methylation patterns. It was hypothesized that the Thuricide, which is made with B. thuringiensis, will not have a significant impact on the methylation patterns of F. ananassa, making it safe for regulated use. DNA from Fragaria ananassa was extracted using a basic extraction method, then exposed to Thuricide for 24, 48, and 72 hours. The exposed DNA went through bisulfite sequencing using the Zymo Research methylation kit, then digested by the hpaii enzyme. The newly digested DNA was analyzed using a fluorometer to determine fluorescence values, which indicated methylation levels. The mean fluorescence values across all groups are unexpectedly similar, with averages close to -0.0056. The control group has the lowest mean fluorescence (-0.0061), while the 48-hour group has the highest (-0.0051). Based on the Kruskal-Wallis test that was ran at an alpha value of ��=.05, the results of this research were sufficient enough to reject the null hypothesis. It was later found in a Post-Hoc Mann-Whitney U Test with Bonferroni Correction that there was a significant difference between the 24 hours and 48 hours. It was concluded that Thuricide did have a slight, but not majorly significant impact on the DNA of F. ananassa, suggesting that it could be used and regulated as a valid replacement for synthetic pesticides in agriculture
The Effect of Different Concentrations of Oyster Shell and Casein Powder on the Thermal Stability of Polyurethane
No full textThe call for environmental reforms has been increasing. prominent in the 21st century. Polyurethane foam, commonly found in buildings and household equipment, poses environmental risks, particularly when burned, as it can contribute to wildfire spread. To address this issue, scientists have explored bio-based flame retardants as protective measures. This project investigated the effectiveness of two bio-based flame retardants—casein and oyster shell—both independently and in combination. The hypothesis suggested that a synergistic combination of oyster shell and casein would prove more effective than either material alone or the control. This was based on the understanding that while higher oyster shell concentrations can decrease effect the tensile strength of polyurethane, combining it with casein could compensate for these effects while maintaining oyster shell\u27s effective char layer formation. The experiment involved creating four intumescent flame retardants: casein alone, oyster shell alone, a combination of both, and a control. Statistical analysis using ANOVA with an α-value of 0.05 revealed significant differences between the experimental groups and the control (p-value \u3c 0.001, α= 0.05). The post-hoc-tukey test, as well at the F=23.0 demonstrated significant differences between experimental groups, except between the combination and casein and casein and oyster. The hypothesis was partially supported, as significant differences were found between most groups, though the combination of casein and oyster shell did not show significant improvement over casein alone but did so over oyster shell alone. This research contributes to the understanding of bio-based flame retardants and their potential applications in environmental protection
The Effect of the Combination of Propolis and Helianthus annuus as an Alternative Ultraviolet Protection
No full textAs the largest organ in the body, the skin is exposed to external factors like bacteria, irritants, and ultraviolet (UV) light, which penetrates its protective barrier. Therefore, it is important to create new photoprotective formulas that help protect the skin barrier. The purpose of this study was to examine the UV-blocking effects of combining propolis and Helianthus annuus (sunflower seed oil) as a natural alternative to commercial sunscreens. Both substances have individual photoprotective properties, but their combined effectiveness has not been extensively studied. It was hypothesized that if propolis and Helianthus annuus were combined, the UV intensity passing through coated the watch glass would decrease due to the photoprotective abilities the oils contain.Using a UV sensor, the amount of UV radiation blocked by watch glasses coated with either propolis, Helianthus annuus, or their combination was measured. Results showed that the combination blocked an average of 24.12% of UV radiation, outperforming propolis (13.53%) and Helianthus annuus (8.48%), but was less effective than commercial sunscreen (65.22%). These results were confirmed with a one way ANOVA test that showed the significant differences between the groups (p \u3c 0.0001). A tukey test was then run to observe the significance in the combination of H. annuus and propolis compared to the extracts separately. It was found that the combination of extracts blocked a higher percentage of UV than they did by themselves. These findings suggest that combining propolis and Helianthus annuus could be a promising natural option for UV protection
The Effects of Propane-1, 2-diol on the Evaporation Rate of Ballpoint Pen Ink
No full textWriting with ink pens has stressed people who constantly worry about getting ink on their skin or clothes. Many studies have shown that ink has solvents that evaporate fairly quickly, leading to ink drying. While this is true in some cases, some left-handed people still deal with the struggle of having to avoid ink. The purpose of this study was to determine if adding propane-1,2 diol to ballpoint pen ink would decrease its drying time. It was hypothesized that adding more propane-1, 2 diol to ink would continue to decrease its drying time. A 50 mL beaker with 10 mL of ballpoint pen ink and a stack of paper was set up to run the experiment. Q-tips were used to dip into the ink and write on the paper. When the Q-tips were lifted from the paper, a stopwatch was set to time how long it took for the ink to dry. After 30 trials were completed, 1 mL of propane-1,2 diol was added to the ink, and 30 more trials were run. After another 30 trials were completed, 3 mL of propane-1, 2 diol were added to the ink, and 30 more trials were completed. The addition of propane-1, 2 diol resulted in no significant trend that suggested the solvent affected the drying time of ink. It was determined that the primary cause of error was experimentation strategy and timing. Precision and better planning would produce better results
Creating a Functional Prototype Passive-‘Powered’ Lower Limb Exoskeleton that Assists in Walking with Heavy Objects
No full textPhysically demanding jobs, such as construction and warehouse work, are often limited by the body’s endurance, leading to fatigue and reduced productivity. While technologies like robotic arms and mechanical claws offer assistance, they are typically expensive, specialized, and require extensive training. Exoskeletons, wearable devices that enhance the body’s movements by applying additional force, provide a more adaptable and affordable alternative. This study aimed to design and evaluate a passive lower limb exoskeleton prototype to reduce physical effort during tasks like lifting or carrying heavy loads. It was hypothesized that a spring-based exoskeleton, storing and releasing energy during joint flexion and extension, would increase the force output of a model leg, reducing potential metabolic costs. Compared to active exoskeletons, passive designs are lighter and more efficient due to the absence of heavy actuators. A model leg simulated human stepping motion, and the force produced was measured with and without the exoskeleton. Results showed that both knee and ankle exoskeletons increased force output, with the knee exoskeleton having a greater impact, supporting its potential to reduce metabolic cost most effectively
The Combined Effects of Myristica fragrans and Curcumin on Saccharomyces cerevisiae serving as a Cancer Cell Model
No full textThe relentless pursuit of understanding and overcoming cancer has shaped one of the most critical scientific endeavors of our time. Many lab studies have been conducted to find new cures using Saccharomyces cerevisiae as a model for cancer cells due to both types of cells preferring glycolysis even in the presence of oxygen. Many studies have shown Myristica fragrans and curcumin to have antifungal properties due to the phytochemicals present in them. The purpose of the study was to assess the combined properties of Myristica fragrans (nutmeg) and curcumin as a potential cure against cancer cells using a model organism, S. cerevisiae. It was hypothesized that as the combined concentration of (nutmeg) and curcumin would increase, the antifungal inhibitory properties against S. cerevisiae would increase, due to the anticancer properties present in both Myristica fragrans and curcumin present in the phytochemicals. In order to test this, three solutions were created with one control of olive oil and two experimental groups with varying concentrations of nutmeg and curcumin. 30 disks were soaked in each solution and placed onto a petri dish that was streaked with yeast. After the 72-hour incubation period, no zone of inhibition was apparent for the control and both experimental groups. It was determined that the primary cause of these results was due to the solutions separating too quickly which altered the amount of phytochemicals saturating the disks. The usage of a better solvent would have allowed for the results to be measurable